SBIR-STTR Award

Triplet repeat expansions of specific DNA loci have recently been shown to underlie ten neurological disorders, including Huntington's disease, fragile X syndrome, and myotonic dystrophy, and are suspected to be involved in several more. Polymerase chain reaction based analysis of the number of repeats provides diagnostic information. There is a need to develop methodologies for automated, high-throughput DNA analysis for genetic typing of disease markers and for linkage analysis Of undiscovered genes. This proposal describes development of integrated reagents and procedures to perform di-, trio and tetra-nucleotide repeat size typing of fluorescent PCR fragments by capillary array electrophoresis. The goal of Phase I is to optimize multiplex genetic typing protocols for a 96-capillary system and demonstrate typing of CA- repeat polymorphisms with 1,800 samples from families affected with hemochromatosis. The integrated methods, instrument, and software will have the capacity to analyze up to 5.5 million genotypes per year, increasing throughput by a factor of nine over current systems. Our goals are to validate the capillary array genetic analysis system for typing of Huntington's disease, fragile X syndrome and myotonic dystrophy, and test a proposed genotype-phenotype correlation of multiple sclerosis progression with repeat polymorphisms in the control region of the myelin basic protein gene.National Institute of Neurological Disorders and Stroke (NINDS)

Our long term goal is to develop methods for automated, high-throughput DNA analysis for genetic typing of disease markers. We propose to develop an integrated instrument system and procedures to perform simple sequence repeat sizing of fluorescent PCR figments by capillary array electrophoresis. During Phase II we propose to: 1) Test and Optimize the integrated 4-color CAE system using protocols developed in Phase I for linkage analysis; 2) Enhance genotyping software by evaluating sizing algorithms and by integrating genotyping and pedigree editing tools; 3) Optimize mutation screening methods using known normal and disease-related alleles. The CAE system performance will then be validated in a molecular genetics testing laboratory. The studies in this proposal will demonstrate the utility and applicability of CAE to a broad range of genetic typing applications. The integrated methods, instrument, and software will have the capacity to analyze up to 9.1 million genotypes per year, increasing throughput by a factor of 5-7 over current systems. Availability of this high-throughput system will significantly enhance the application of genetic typing in the health sciences.Proposed commercial application:A capillary array genetic analysis system has significant potential to improve the U.S. economic growth by accelerating the rate that new genetic information is generated. Reliable, cost-efficient genetic typing will have significant impact on healthcare, agribusiness, forensic analysis, and pharmaceutical industry sectors. The systems offer the potential to significantly control healthcare costs through economical yet accurate early diagnosis leading to prevention or effective treatment of serious genetic disorders, including neurodegenerative and neuropsychiatric illnesses.Thesaurus termsbiomedical equipment development, capillary electrophoresis, diagnosis design /evaluation, genetic disorder diagnosis, nucleic acid repetitive sequence DNA, allele, computer assisted sequence analysis, genetic marker, genotype, neurogenetics computer program /software, fluorescence, human genetic material tag, linkage mapping, polymerase chain reactionNational Institute of Neurological Disorders and Stroke (NINDS)